The confocal Core Family will provide equipment and technical expertise for co-investigators in cystic fibrosis research at the Mayo Foundation. This unique core facility will allow investigators to examine subcellular localization of proteins and interacting proteins, to determine levels and time course or changes in intracellular ions and other fluorescently- labeled second messengers, to use caged compounds to examine the physiologic mechanism of rapid changes in intracellular second messengers, to routinely make three-dimensional reconstructions of living and fixed cells at high resolution, and to make use of optical tweezer technology to manipulate intracellular organelles. This confocal core facility is a state-of-the-art facility with several unique capabilities, including the ability to use two-photon excitation for imaging and high resolution photolysis of caged compounds. In the near future, technical development of a new spectral imaging system will be completed, allowing up to eight fluorescent dyes of various wavelengths to be imaged simultaneously. In addition, the confocal facility has implemented infrared optical tweezers, and has extensive three-dimensional reconstruction software capabilities. In particular, the core facility will provide assistance in the imaging of improperly folded and mutant CFTR gene products, proteins associated with pI/Cln and their intracellular distribution, and the co-localization of mucin with other proteins and its expression within tissues and cells.